Detection and Quantification with 16S rRNA Probes of Planktonic Methylotrophic Bacteria in a Floodplain Lake

Abstract Two approaches employing 16S rRNA oligonucleotide probes, in situ hybridization combined with ³³P-autoradiography and ³²P-labeled slot-blot hybridizations on nitrocellulose filters, were used to enumerate methylotrophic bacteria in the water column of Ryans 1 Billabong, a small floodplain lake in northeastern Victoria, Australia. Methylotrophic bacterioplankton numbered 0.6–1.2 × 10⁹ cells liter⁻¹ in the winter of 1994, and 0.8–5.5 × 10⁹ cells liter⁻¹ in the summer of 1994–95. This was equivalent to 10–46% of total bacterioplankton cell counts, determined via epifluorescence microscopy. Methylotrophic bacteria were not detected in the water column of the nearby Kiewa River, and a set of laboratory controls indicated that the high abundance of methylotrophs in the billabong samples was not a methodological artifact. There was no change, with water depth, in total bacterioplankton or methylotroph abundance in winter, a result consistent with the water column being well mixed at this time of year (dissolved O₂ concentrations 5–7 mg liter⁻¹; dissolved methane concentrations <60 μg liter⁻¹, or <5% methane saturation, at all depths). In summer the billabong became diurnally stratified (dissolved O₂ concentrations <2 mg liter⁻¹ at water depths of >45 cm; dissolved methane concentrations <100 μg liter⁻¹ at the surface, but >500 μg liter⁻¹ near the sediments) and there was a correspondingly marked increase in the abundance of total bacterioplankton and methylotrophs with depth. In situ hybridizations and slot-blot hybridizations both indicated that type II methylotrophs (detected with a probe specific for the 9-α subgroup of Proteobacteria) were markedly less abundant than were type I and X methylotrophs (detected with a probe specific for the 10-γ subgroup of Proteobacteria). Received: 12 March 1996; Accepted: 2 October 1996     

Bacterial assemblages in rivers and billabongs of Southeastern Australia

Billabongs, lentic waterbodies common to the floodplain of Australian rivers, differ considerably from the lotic riverine environment in terms of hydrology, physiochemical characteristics, and biological assemblages present. As little is known regarding the bacterial ecology of billabong habitats, a comparison was made of the bacterial assemblages in the water column of seven paired river/billabong sites in the Murray-Darling Basin of southeastern Australia. Billabongs supported larger populations of bacteria (1–157×10⁹ cells liter⁻¹; 11–10,270 μg C liter⁻¹) than did rivers (1–10×10⁹ cells liter⁻¹; 6–143 μg C liter⁻¹). Phospholipid analyses confirmed that billabongs (14–111 μg phospholipid fatty acid liter⁻¹) had larger bacterial populations than rivers (<12 μg liter⁻¹). Bacterial production, measured with³H-leucine, was also greater in billabongs (0.28–3.05 μg C liter⁻¹ hour⁻¹) than rivers (0.05–0.62 μg C liter⁻¹ hour⁻¹). Production calculated from the frequency of dividing cells confirmed this conclusion, and suggested bacterial production in some billabongs could exceed 100 μg C liter⁻¹ hour⁻¹. An INT-formazan method indicated that usually <25% of bacterial cells were active in either habitat, but this was probably an underestimate of the bona fide value. Turnover times of glucose were usually shorter in billabongs, and the cell-specific activity greater for billabong than river assemblages. The factors most likely to be responsible for the differences between the bacterial assemblages in rivers and billabongs relate to hydrological regime and the availability of organic carbon substrates.     

Detrital Controls on Soil Solution N and Dissolved Organic Matter in Soils: A Field Experiment

We established a long-term field study in an old growth coniferous forest at the H.J. Andrews Experimental Forest, OR, USA, to address how detrital quality and quantity control soil organic matter accumulation and stabilization. The Detritus Input and Removal Treatments (DIRT) plots consist of treatments that double leaf litter, double woody debris inputs, exclude litter inputs, or remove root inputs via trenching. We measured changes in soil solution chemistry with depth, and conducted long-term incubations of bulk soils from different treatments in order to elucidate effects of detrital inputs on the relative amounts and lability of different soil C pools. In the field, the addition of woody debris increased dissolved organic carbon (DOC) concentrations in O-horizon leachate and at 30 cm, but not at 100 cm, compared to control plots, suggesting increased rates of DOC retention with added woody debris. DOC concentrations decreased through the soil profile in all plots to a greater degree than did dissolved organic nitrogen (DON), most likely due to preferential sorption of high C:N hydrophobic dissolved organic matter (DOM) in upper horizons; percent hydrophobic DOM decreased significantly with depth, and hydrophilic DOM had a much lower and less variable C:N ratio. Although laboratory extracts of different litter types showed differences in DOM chemistry, percent hydrophobic DOM did not differ among soil solutions from different detrital treatments in the field, suggesting that microbial processing of DOM leachate in the field consumed easily degradable components, thus equalizing leachate chemistry among treatments. Total dissolved N leaching from plots with intact roots was very low (0.17 g m⁻² year⁻¹), slightly less than measured deposition to this very unpolluted forest (~s 0.2 g m⁻² year⁻¹). Total dissolved N losses showed significant increases in the two treatments without roots whereas concentrations of DOC decreased. In these plots, N losses were less than half of estimated plant uptake, suggesting that other mechanisms, such as increased microbial immobilization of N, accounted for retention of N in deep soils. In long-term laboratory incubations, soils from plots that had both above- and below-ground litter inputs excluded for 5 years showed a trend towards lower DOC loss rates, but not lower respiration rates. Soils from plots with added wood had similar respiration and DOC loss rates as control soils, suggesting that the additional DOC sorption observed in the field in these soils was stabilized in the soil and not readily lost upon incubation.     

Factors Controlling the Year-Round Variability in Carbon Flux Through Bacteria in a Coastal Marine System

Data from several years of monthly samplings are combined with a 1-year detailed study of carbon flux through bacteria at a NW Mediterranean coastal site to delineate the bacterial role in carbon use and to assess whether environmental factors or bacterial assemblage composition affected the in situ rates of bacterial carbon processing. Leucine (Leu) uptake rates [as an estimate of bacterial heterotrophic production (BHP)] showed high interannual variability but, on average, lower values were found in winter (around 50 pM Leu⁻¹ h⁻¹) as compared to summer (around 150 pM Leu⁻¹ h⁻¹). Leu-to-carbon conversion factors ranged from 0.9 to 3.6 kgC mol Leu⁻¹, with generally higher values in winter. Leu uptake was only weakly correlated to temperature, and over a full-year cycle (in 2003), Leu uptake peaked concomitantly with winter chlorophyll a (Chl a) maxima, and in periods of high ectoenzyme activities in spring and summer. This suggests that both low molecular weight dissolved organic matter (DOM) released by phytoplankton, and high molecular weight DOM in periods of low Chl a, can enhance BHP. Bacterial respiration (BR, range 7–48 μg C l⁻¹ d⁻¹) was not correlated to BHP or temperature, but was significantly correlated to DOC concentration. Total bacterial carbon demand (BHP plus BR) was only met by dissolved organic carbon produced by phytoplankton during the winter period. We measured bacterial growth efficiencies by the short-term and the long-term methods and they ranged from 3 to 42%, increasing during the phytoplankton blooms in winter (during the Chl a peaks), and in spring. Changes in bacterioplankton assemblage structure (as depicted by denaturing gradient gel electrophoresis fingerprinting) were not coupled to changes in ecosystem functioning, at least in bacterial carbon use.     

Quantitative and Qualitative Aspects of Dissolved Organic Carbon Leached from Senescent Plants in an Oligotrophic Wetland

We conducted a series of experiments whereby dissolved organic matter (DOM) was leached from various wetland and estuarine plants, namely sawgrass (Cladium jamaicense), spikerush (Eleocharis cellulosa), red mangrove (Rhizophora mangle), cattail (Typha domingensis), periphyton (dry and wet mat), and a seagrass (turtle grass; Thalassia testudinum). All are abundant in the Florida Coastal Everglades (FCE) except for cattail, but this species has a potential to proliferate in this environment. Senescent plant samples were immersed into ultrapure water with and without addition of 0.1% NaN₃ (w/ and w/o NaN₃, respectively) for 36 days. We replaced the water every 3 days. The amount of dissolved organic carbon (DOC), sugars, and phenols in the leachates were analyzed. The contribution of plant leachates to the ultrafiltered high molecular weight fraction of DOM (>1 kDa; UDOM) in natural waters in the FCE was also investigated. UDOM in plant leachates was obtained by tangential flow ultrafiltration and its carbon and phenolic compound compositions were analyzed using solid state ¹³C cross-polarization magic angle spinning nuclear magnetic resonance (¹³C CPMAS NMR) spectroscopy and thermochemolysis in the presence of tetramethylammonium hydroxide (TMAH thermochemolysis), respectively. The maximum yield of DOC leached from plants over the 36-day incubations ranged from 13.0 to 55.2 g C kg⁻¹ dry weight. This amount was lower in w/o NaN₃ treatments (more DOC was consumed by microbes than produced) except for periphyton. During the first 2 weeks of the 5 week incubation period, 60–85% of the total amount of DOC was leached, and exponential decay models fit the leaching rates except for periphyton w/o NaN₃. Leached DOC (w/ NaN₃) contained different concentrations of sugars and phenols depending on the plant types (1.09–7.22 and 0.38–12.4 g C kg⁻¹ dry weight, respectively), and those biomolecules comprised 8–34% and 4–28% of the total DOC, respectively. This result shows that polyphenols that readily leach from senescent plants can be an important source of chromophoric DOM (CDOM) in wetland environments. The O-alkyl C was found to be the major C form (55±9%) of UDOM in plant leachates as determined by ¹³C CPMAS NMR. The relative abundance of alkyl C and carbonyl C was consistently lower in plant-leached UDOM than that in natural water UDOM in the FCE, which suggests that these constituents increase in relative abundance during diagenetic processing. TMAH thermochemolysis analysis revealed that the phenolic composition was different among the UDOM leached from different plants, and was expected to serve as a source indicator of UDOM in natural water. Polyphenols are, however, very reactive and photosensitive in aquatic environments, and thus may loose their plant-specific molecular characteristics shortly. Our study suggests that variations in vegetative cover across a wetland landscape will affect the quantity and quality of DOM leached into the water, and such differences in DOM characteristics may affect other biogeochemical processes.     

Characterisation and reactivity continuum of dissolved organic matter in forested headwater catchments of Andean Patagonia

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A Seasonal Study of Bacterial Community Succession in a Temperate Backwater System,Indicated by Variation in Morphotype Numbers,Biomass, and Secondary Production

Abstract The investigation of the bacterial community in the Kühw?rter Wasser, a macrophyte-dominated arm of the River Danube backwater system near Vienna, revealed that variation in microbial densities and biomass could be related to a characteristic sequence in morphotype composition over the seasons. Maximal bacterial cell numbers and biomass occured in early summer, with values of up to 9 × 10⁹ cells l⁻¹ and 122 μg C l⁻¹, respectively, caused by a massive increase of vibrio-shaped cells. On the other hand, in early spring, filamentous bacteria were responsible for a marked increase in bacterial biomass, making up 40% of the total bacterial biomass. Over the year, rod-shaped cells were the dominating morphotype, while the biomass of cocci was rather negligible. In winter, cell numbers and biomass showed minimal values with 2.0 × 10⁹ cells l⁻¹ and 28 μg C l⁻¹, respectively, and bacteria were considered to be substrate and temperature limited during this period. Saturation values of the incorporation of ³H-thymidine into DNA, for the estimation of bacterial secondary production, varied seasonally, ranging from 5 nm to 40 nm. Thus, saturation experiments needed to be conducted on a regular basis. Also, the amount of labeled thymidine in the DNA, as a percentage of labeled thymidine in the TCA precipitate, varied over the year. Minimum values of 45% were recorded during the cold season, while maximum values of 75–80% at the beginning of June coincided with high chlorophyll a values and minimal K _m-values derived from saturation experiments. The potential role of the nitrogen-rich nucleoside thymidine as a readily utilizable substrate for bacteria during labeling experiments, under varying conditions of substrate availability, is discussed. Bacterial secondary production rates ranged from 0.3 μg C l⁻¹ h⁻¹ in winter to values of 10 μg C l⁻¹ h⁻¹ in August, where phytoplanktonic biomass reached the summer maximum, and bacterial biomass was calculated to be renewed 3 times per day. An estimation of the bacterial carbon demand showed that for the major part of the year, with the exception of early spring, the bacterioplankton community in the Kühw?rter Wasser was dependent on carbon sources other than phytoplanktonic primary production. Received: 22 March 1996; Revised: 1 August 1996     

Distribution and activity of bacteria in deep granitic groundwaters of southeastern sweden

This study investigated the distribution of bacteria in groundwater from 16 different levels in five boreholes in granite bedrock down to a maximum of 860 m. Enrichment cultures were used to assay the groups of bacteria present. Autoradiographic studies with¹⁴C- or³H-labeled formate, methanol, acetate, lactate, glucose, sodium bicarbonate, leucine, glutamine, thymidine, orN-acetyl-glucosamine were used to obtain information about bacteria active in substrate uptake. The biofilm formation potential was studied in one borehole. The chemical environment in the groundwater was anaerobic with an E_h between −112 and −383 mV, a pH usually around 8, and a temperature range of 10.2 to 20.5°C, depending on the depth. The organic content ranged between <0.5 and 9.5 mg total organic carbon liter⁻¹. Carbon dioxide, hydrogen, hydrogen sulfide, and methane were present in the water. The nitrate, nitrite, and phosphate concentrations were close to, or below, the detection limits, while there were detectable amounts of NH ₄ ⁺ in the range of 4 to 330 μg liter⁻¹. The average total number of bacteria was 2.6×10⁵ bacteria ml⁻¹, as determined with an acridine organge direct-count (AODC) technique. The average number of bacteria that grew on a medium with 1.5 g liter⁻¹ of organic substrate was 7.7×10³ colony-forming units (CFU) ml⁻¹. The majority of these were facultatively anaerobic, gram-negative, nonfermenting heterotrophs. Enrichment cultures indicated the presence of anaerobic bacteria capable of growth on C-1 compounds and hydrogen, presumably methanogenic bacteria. Most probable number assays with sulfate and lactate revealed up to 5.6×10⁴ viable sulfate-reducing bacteria per ml. A biofilm development experiment indicated an active attached microbial population. Active substrate uptake could not be registered with the bulk water populations, except for an uptake of leucine not associated with growth. The bulk water microbial cells in deep groundwater may be inactive cells detached from active biofilms on the rock surface.     

Balance between primary and bacterial production in North Patagonian shallow lakes

The shallow Andean North Patagonian lakes are suitable environments for the evaluation of autotrophic and heterotrophic production under a scenario of high radiation in high dissolved organic matter (DOM) systems. We aimed to study the balance between primary and bacterial production in three shallow Andean lakes, in a summer sampling (high irradiance condition). Our hypothesis is that two factors would interact: high light and high DOM, affecting bacteria and algae. We carried out experiments of bacterial production (BP) by measuring [¹⁴C]-l-leucine incorporation and PP by ¹⁴C uptake in two fractions: picophytoplankton and phytoplankton >2 μm. Cell abundance, chlorophyll a, nutrients, DOM, light, and temperature were also measured. The contribution of picophytoplankton to total primary production (PP) was, in general, very high exceeding 50%. Picophytoplankton was photosynthetically more efficient than the larger autotrophs in all lakes. We observed a decrease in PP at surface levels due the high solar irradiances, while BP was not affected. Changes in the PP:BP ratios were observed in relation to DOM content and light effect. Our data indicate that the amount of available DOM drives the balance between PP and BP. However, solar radiation should be included as an important factor since PP:BP ratio may decrease because of PP photoinhibition.     

Bacterial Contribution to Dissolved Organic Matter in Eutrophic Lake Kasumigaura,Japan

Incubation experiments using filtered waters from Lake Kasumigaura were conducted to examine bacterial contribution to a dissolved organic carbon (DOC) pool. Bacterial abundance, bacterial production, concentrations of DOC, total dissolved amino acids (TDAA), and total dissolved neutral sugars (TDNS) were monitored during the experiments. Bacterial production during the first few days was very high (20 to 35 μg C liter⁻¹ day⁻¹), accounting for 40 to 70% of primary production. The total bacterial production accounted for 34 to 55% of the DOC loss during the experiment, indicating high bacterial activities in Lake Kasumigaura. The DOC degradation was only 12 to 15%, whereas the degradation of TDAA and TDNS ranged from 30 to 50%, suggesting the preferential usage of TDAA and TDNS. The contribution of bacterially derived carbon to a DOC pool in Lake Kasumigaura was estimated using d-amino acids as bacterial biomarkers and accounted for 30 to 50% of the lake DOC. These values were much higher than those estimated for the open ocean (20 to 30%). The ratio of bacterially derived carbon to bulk carbon increased slightly with time, suggesting that the bacterially derived carbon is more resistant to microbial degradation than bulk carbon. This is the first study to estimate the bacterial contribution to a DOC pool in freshwater environments. These results indicate that bacteria play even more important roles in carbon cycles in freshwater environments than in open oceans and also suggests that recent increases in recalcitrant DOC in various lakes could be attributed to bacterially derived carbon. The potential differences in bacterial contributions to dissolved organic matter (DOM) between freshwater and marine environments are discussed.     

Simultaneous utilization of glucose and D-xylose by Candida shehatae in a chemostat

The kinetics of biomass formation, D-xylose utilization, and mixed substrate utilization were determined in a chemostat using the yeast Candida shehatae. The maximum growth rate of C. shehatae grown aerobically on D-xylose was 0.42 h⁻¹ and the Monod constant, K _s, was 0.06 g L⁻¹. The biomass yield, Y _{X/S}, ranged from 0.40 to 0.50 g g⁻¹ over a dilution rate range of 0.2–0.3 h⁻¹, when C. shehatae was grown on pure D-xylose. Mixtures of D-xylose and glucose (∼1 : 1) were simultaneously utilized over a dilution rate from 0.15 to 0.35 h⁻¹ at pH 3.5 and 4.5, but pH 3.5 reduced μ_max and reduced the dilution rate range over which D-xylose was utilized in the presence of glucose. At pH 4.5, μ_max was not reduced with the mixed sugar feed and the overall or lumped K _s value was not significantly increased (0.058 g L⁻¹ vs 0.06 g L⁻¹), when compared to a pure D-xylose feed. Kinetic data indicate that C. shehatae is an excellent candidate for chemostat production of value added products from renewable carbon sources, since simultaneous mixed substrate utilization was observed over a wide range of growth rates on a 1 : 1 mixture of glucose and D-xylose. Received 21 August 1997/ Accepted in revised form 28 May 1998     

Sugarcane Straw and its Phytochemicals as Growth Regulators of Weed and Crop Plants

The present study investigates the effect of soil amended with sugarcane straw leachate and its constituents on root elongation of weed and crop plants. The influence of soil biotic and abiotic factors on plant growth were also evaluated through assays in both autoclaved soil and sand. In unsterile soil, straw leachates stimulated root growth of some test plants at 6 g dry straw ls⁻¹ and inhibited root elongation at higher concentrations. A bioassay guided process of the bioactive leachate constituents led to the isolation of vanillic, syringic and ferulic acids. These compounds were also assayed on the test plants in the three mentioned substrates. In unsterile soil, phenolics stimulated the growth of some species at 19 mg l⁻¹. Higher phenolics concentrations were inhibitory. The concentration needed to inhibit 25% root elongation (EC₂₅) was calculated from the dose–response curves. The concentration of phenolics in the leachate (64 g dry straw l⁻¹) was estimated to be 187 mg l⁻¹ (ferulic acid), 131 mg l⁻¹ (vanillic acid) and 20 mg l⁻¹ (Syringic acid). In general, these concentrations were below the EC₂₅ values determined in unsterile soil indicating that these compounds cannot completely explain the strong inhibitory activity of sugarcane straw leachates. The role of soil factors on phytotoxicity of sugarcane straw leachate and its identified growth regulators is also discussed.     

The mycobiota of landfill leachates in the pretreatment process in a sequencing batch reactor

The paper discusses the dynamics of the accumulation of microscopic fungi, depending on the sludge load (Bx), in activated sludge used for landfill leachate pretreatment. The propagule washout from the sludge into pretreated leachates is determined, including genera and species that may threaten environmental health. An increased accumulation of microscopic fungi in sludge flocs occurred at Bx=0.23−0.45 mg chemical oxygen demand (COD) mg⁻¹ d⁻¹. Microscopic fungi were eluted at the maximal Bx value tested of 1.64 mg COD mg⁻¹ d⁻¹. Both the activated sludge and the leachate runoff from the sequencing batch reactor (SBR) pose health risks to the environment due to the occurrence of fungi such as Aspergillus fumigatus, Purpureocillium lilacinum, Cyberlindnera jadinii (C. utilis), Geotrichum candidum and G. fragrans. Their count is sufficient to cause multi-organ infections in homeothermal animals and in humans.     

Bacterial Colonization and Ectoenzymatic Activity in Phytoplankton-Derived Model Particles. Part II. Cleavage and Uptake of Carbohydrates

Abstract The bacterial colonization and development of the ectoenzymatic glucosidase activity and glucose uptake were followed together with bacterial growth (measured as thymidine incorporation) in laboratory experiments, using phytoplankton-derived particles incubated in rolling tanks. Bacterial colonization of the particles was rapid. In the particles, bacterial turnover rates (production/biomass) were low (0.02 to 0.14 d⁻¹). In the ambient water, turnover rates increased from 0.1 d⁻¹ to 23.3 d⁻¹, until the end of the experiment. In the control, lacking any particles, turnover of bacteria ranged from 0.3 to 7.6 d⁻¹. Similarly, glucose uptake rates, per bacterium, were 1 to 2 orders of magnitude lower for particle-attached bacteria than for their free-living counterparts. Generally, K_m values for glucosidase activity declined, over the incubation period, in particles and free-living bacteria until 168 h, and slightly increased, thereafter, to values of approximately 0.1 μM. Particle-attached bacteria exhibited significantly lower uptake rates of both thymidine and glucose, per bacterium, throughout the incubation. The per-cell ectoenzymatic activity was similar in particle-associated and free-living bacteria during the initial phase of the experiment, but was significantly higher after ≈200 h. Dissolved total (TCHO), as well as monomeric carbohydrates (MCHO), declined continuously in both particles and ambient water; they remained constant in the control; TCHO comprised about 50% of the dissolved organic carbon (DOC) in the particles. In ambient water TCHO contribution to DOC varied, with only one exception, between 25 and 45%; and in the control, between 20 and 50%. The shift detectable in the relation between ectoenzymatic activity and uptake of glucose between free-living and attached bacteria over the incubation period may reflect changes in the physiological status of the bacteria. Received: 3 February 1997; Accepted: 6 November 1997     

Enhanced l-lysine production in threonine-limited continuous culture of Corynebacterium glutamicum by using gluconate as a secondary carbon source with glucose

In order to improve the production rate of l-lysine, a mutant of Corynebacterium glutamicum ATCC 21513 was cultivated in complex medium with gluconate and glucose as mixed carbon sources. In a batch culture, this strain was found to consume gluconate and glucose simultaneously. In continuous culture at dilution rates ranging from 0.2 h⁻¹ to 0.25 h⁻¹, the specific l-lysine production rate increased to 0.12 g g⁻¹ h⁻¹ from 0.1 g g⁻¹ h⁻¹, the rate obtained with glucose as the sole carbon source [Lee et al. (1995) Appl Microbiol Biotechnol 43:1019–1027]. It is notable that l-lysine production was observed at higher dilution rates than 0.4 h⁻¹, which was not observed when glucose was the sole carbon source. The positive effect of gluconate was confirmed in the shift of the carbon source from glucose to gluconate. The metabolic transition, which has been characterized by decreased l-lysine production at the higher glucose uptake rates, was not observed when gluconate was added. These results demonstrate that the utilization of gluconate as a secondary carbon source improves the maximum l-lysine production rate in the threonine-limited continuous culture, probably by relieving the limiting factors in the lysine synthesis rate such as NADPH supply and/or phosphoenolpyruvate availability. Received: 16 May 1997 / Received revision: 28 August 1997 / Accepted: 29 August 1997     

Environmental Dissolved Organic Matter Governs Biofilm Formation and Subsequent Linuron Degradation Activity of a Linuron-Degrading Bacterial Consortium

It was examined whether biofilm growth on dissolved organic matter (DOM) of a three-species consortium whose members synergistically degrade the phenylurea herbicide linuron affected the consortium''s integrity and subsequent linuron-degrading functionality. Citrate as a model DOM and three environmental DOM (eDOM) formulations of different quality were used. Biofilms developed with all DOM formulations, and the three species were retained in the biofilm. However, biofilm biomass, species composition, architecture, and colocalization of member strains depended on DOM and its biodegradability. To assess the linuron-degrading functionality, biofilms were subsequently irrigated with linuron at 10 mg liter⁻¹ or 100 μg liter⁻¹. Instant linuron degradation, the time needed to attain maximal linuron degradation, and hence the total amount of linuron removed depended on both the DOM used for growth and the linuron concentration. At 10 mg liter⁻¹, the final linuron degradation efficiency was as high as previously observed without DOM except for biofilms fed with humic acids which did not degrade linuron. At 100 μg liter⁻¹ linuron, DOM-grown biofilms degraded linuron less efficiently than biofilms receiving 10 mg liter⁻¹ linuron. The amount of linuron removed was more correlated with biofilm species composition than with biomass or structure. Based on visual observations, colocalization of consortium members in biofilms after the DOM feed appears essential for instant linuron-degrading activity and might explain the differences in overall linuron degradation. The data show that DOM quality determines biofilm structure and composition of the pesticide-degrading consortium in periods with DOM as the main carbon source and can affect subsequent pesticide-degrading activity, especially at micropollutant concentrations.     

In situ release of coral mucus by <Emphasis Type="Italic">Acropora</Emphasis> and its influence on the heterotrophic bacteria

In situ mucus release by Acropora nobilis and degradation of mucus from A. nobilis and Acropora formosa, by heterotrophic bacteria were investigated at Bidong and Tioman Island, Malaysia. Mucus release rate for A. nobilis was on average 38.7 ± 35.2 mg C m⁻² h⁻¹, of which ca. 70% consisted of dissolved organic carbon (DOC) and 30% particulate organic carbon (POC). In the mucus degradation experiment, seawater-mucus mixtures were incubated and compared with control runs for 24 h. Bacterial abundance in the seawater-mucus mixture increased significantly and coincided with a decline in DOC concentration. In controls, bacteria and DOC did not significantly change. The coral mucus had a high content of inorganic phosphate. It is suggested that the coral mucus rich in DOC and phosphate can induce the high bacterial growth.     

Bacterial production in a mesohumic lake estimated from [14C]leucine incorporation rate

Incorporation of [¹⁴C]leucine into proteins of bacteria was studied in a temperate mesohumic lake. The maximum incorporation of [¹⁴C] leucine was reached at a concentration of 30 nm determined in dilution cultures. Growth experiments were used to estimate factors for converting leucine incorporation to bacterial cell numbers or biomass. The initially high conversion factors calculated by the derivative method decreased to lower values after the bacteria started to grow. Average conversion factors were 7.09 × 10¹⁶ cells mol^–1 and 7.71 × 10¹⁵ m³ mol^–1, if the high initial values were excluded. Using the cumulative method, the average conversion factor was 5.38 × 10¹⁵ m^–3 mol^–1 I . The empirically measured factor converting bacterial biomass to carbon was 0.36 pg C m^–3 or 33.1 fg C cell^–1. Bacterial production was highest during the growing season, ranging between 1.8 and 13.2 g C liter^–1 day^–1, and lowest in winter, at 0.2–2.9 g C liter^–1 day^–1. Bacterial production showed clear response to changes in the phytoplankton production, which indicates that photosynthetically produced dissolved compounds were used by bacteria. In the epilimnion bacterial production was, on average, 19–33% of primary production. Assuming 50% growth efficiency for bacteria, the allochthonous organic carbon could have also been an additional energy and carbon source for bacteria, especially in autumn and winter. In winter, a strong relationship was found between temperature and bacterial production. The measuring of [¹⁴C]leucine incorporation proved to be a simple and useful method for estimating bacterial production in humic water. However, an appropriate amount of [¹⁴C]leucine has to be used to ensure the maximum uptake of label and to minimize isotope dilution.     

Bacterial Standing Stock, Activity, and Carbon Production during Formation and Growth of Sea Ice in the Weddell Sea, Antarctica

Bacterial response to formation and growth of sea ice was investigated during autumn in the northeastern Weddell Sea. Changes in standing stock, activity, and carbon production of bacteria were determined in successive stages of ice development. During initial ice formation, concentrations of bacterial cells, in the order of 1 × 10⁸ to 3 × 10⁸ liter^-1, were not enhanced within the ice matrix. This suggests that physical enrichment of bacteria by ice crystals is not effective. Due to low concentrations of phytoplankton in the water column during freezing, incorporation of bacteria into newly formed ice via attachment to algal cells or aggregates was not recorded in this study. As soon as the ice had formed, the general metabolic activity of bacterial populations was strongly suppressed. Furthermore, the ratio of [³H]leucine incorporation into proteins to [³H]thymidine incorporation into DNA changed during ice growth. In thick pack ice, bacterial activity recovered and growth rates up to 0.6 day^-1 indicated actively dividing populations. However, biomass-specific utilization of organic compounds remained lower than in open water. Bacterial concentrations of up to 2.8 × 10⁹ cells liter^-1 along with considerably enlarged cell volumes accumulated within thick pack ice, suggesting reduced mortality rates of bacteria within the small brine pores. In the course of ice development, bacterial carbon production increased from about 0.01 to 0.4 μg of C liter^-1 h^-1. In thick ice, bacterial secondary production exceeded primary production of microalgae.     

Effect of dissolved organic carbon quality on microbial decomposition and nitrification rates in stream sediments

1. Microbial decomposition of dissolved organic carbon (DOC) contributes to overall stream metabolism and can influence many processes in the nitrogen cycle, including nitrification. Little is known, however, about the relative decomposition rates of different DOC sources and their subsequent effect on nitrification. 2. In this study, labile fraction and overall microbial decomposition of DOC were measured for leaf leachates from 18 temperate forest tree species. Between 61 and 82% (mean, 75%) of the DOC was metabolized in 24 days. Significant differences among leachates were found for labile fraction rates (P < 0.0001) but not for overall rates (P=0.088). 3. Nitrification rates in stream sediments were determined after addition of 10 mg C L^–1 of each leachate. Nitrification rates ranged from below detection to 0.49 μg N mL sediment^–1 day^–1 and were significantly correlated with two independent measures of leachate DOC quality, overall microbial decomposition rate (r=–0.594, P=0.0093) and specific ultraviolet absorbance (r=0.469, P=0.0497). Both correlations suggest that nitrification rates were lower in the presence of higher quality carbon. 4. Nitrification rates in sediments also were measured after additions of four leachates and glucose at three carbon concentrations (10, 30, and 50 mg C L^–1). For all carbon sources, nitrification rates decreased as carbon concentration increased. Glucose and white pine leachate most strongly depressed nitrification. Glucose likely increased the metabolism of heterotrophic bacteria, which then out‐competed nitrifying bacteria for NH₄⁺. White pine leachate probably increased heterotrophic metabolism and directly inhibited nitrification by allelopathy.